Univ Franche-Comte, F-25000 Besancon, France; EA 3181, FED4234, CIC-BT 506, F-25000 Besancon, France.
J Virol Methods. 2013 Nov;193(2):498-502. doi: 10.1016/j.jviromet.2013.07.023. Epub 2013 Jul 24.
HPV 16 and HPV 18 are responsible for more than 75% of cervical cancers and high HPV 16 loads are associated with both prevalent and incident lesions. The objective of the present study was to develop a method allowing the detection and quantitation of HPV 16 and 18 DNA to improve future strategies for cervical cancer screening. A duplex real-time PCR allowing the simultaneous quantitation of both HPV 16 and HPV 18 was carried out. Mixes of HPV 16 and HPV 18 whole genome plasmids were prepared to test a wide range of viral DNA concentrations. The values obtained for each mix of plasmids with the simplex and the duplex PCR were very close to the theoretical values except when a HPV type represented only 1:1000 genome equivalent or lower than the concurrent type. Cervical samples harboring HPV 16, HPV 18 or both types were tested by comparing the results with simplex and duplex real-time PCR assays. HPV 16 and HPV 18 genome titers were similar with the two assays. In conclusion, the real-time duplex PCR proved to be robust for HPV 16 and HPV 18 DNA quantitation.
HPV 16 和 HPV 18 导致超过 75%的宫颈癌，且高 HPV 16 负荷与现患和新发病变均相关。本研究的目的是开发一种可同时检测和定量 HPV 16 和 18 DNA 的方法，以改进未来的宫颈癌筛查策略。本研究进行了一种可同时定量 HPV 16 和 HPV 18 的双实时 PCR。制备 HPV 16 和 HPV 18 全基因组质粒混合物，以测试广泛的病毒 DNA 浓度。用 simplex 和 duplex PCR 检测每个质粒混合物的结果与理论值非常接近，除了 HPV 类型仅为 1:1000 基因组当量或低于共存类型的情况。通过比较 simplex 和 duplex 实时 PCR 检测，对含有 HPV 16、HPV 18 或两种类型的宫颈样本进行了检测。两种检测方法的 HPV 16 和 HPV 18 基因组滴度相似。总之，双实时 PCR 可稳健地定量 HPV 16 和 HPV 18 DNA。
